Molecular cloning of the Shv-1 beta-lactamase gene and construction of an Shv-1 hybridization probe.

We have cloned the Shv-1 beta-lactamase gene from the R1010 plasmid into pACYC184. By subcloning and transposon mutagenesis we have localized the gene to a 1.6 kb BscI-SalI fragment of R1010, which is present in the recombinant plasmid pUB8. A 900 bp PstI fragment of pUB8 was shown to be a specific hybridization probe by testing against plasmids which encode 17 different beta-lactamase enzymes. A comparison was made of the sensitivity of the Shv-1 probe labelled with either [35S]dCTP or with photobiotin.